Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human, Mouse, Rat

Cited:

Rat

Predicted:

Bovine (95%), Canine (93%), Hamster (95%), Porcine (94%). Backed by our 100% Guarantee.

Applications

Validated:

Knockout Validated, Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence

Cited:

Immunohistochemistry-Paraffin, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all HSP27 Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to an internal region of the human Hsp27 protein (within residues 80-150). [Swiss-Prot# P04792]

Reactivity Notes

Immunogen displays the following percentage of sequence identity for non-tested species: Chicken (80%).

Localization

Cytoplasm and nucleus. Translocates to the nucleus during heat shock and resides in sub-nuclear structures known as SC35 speckles or nuclear splicing speckles.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Description

Novus Biologicals Knockout (KO) Validated Rabbit HSP27 Antibody - BSA Free (NBP1-75477) is a polyclonal antibody validated for use in IHC, WB and ICC/IF. Anti-HSP27 Antibody: Cited in 2 publications. All Novus Biologicals antibodies are covered by our 100% guarantee.

Scientific Data Images for HSP27 Antibody - BSA Free

Knockout Validated: HSP27 Antibody - BSA Free [NBP1-75477]

Western Blot: HSP27 Antibody - BSA Free [NBP1-75477]

Western Blot: HSP27 Antibody [NBP1-75477] - Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and HSP27 knockout (KO) HeLa cell line. PVDF membrane was probed with 2 ug/ml of Rabbit Anti-Human HSP27 Polyclonal Antibody (Catalog # NBP1-75477) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (catalog number HAF008). Specific band was detected for HSP27 at approximately 27 kDa (as indicated) in the parental HeLa cell line, but is not detectable in the knockout HeLa cell line. This experiment was conducted under reducing conditions.
Western Blot: HSP27 AntibodyBSA Free [NBP1-75477]

Western Blot: HSP27 AntibodyBSA Free [NBP1-75477]

Western Blot: HSP27 Antibody [NBP1-75477] - Analysis of Jurkat cell lysate (30ug) using anti-Hsp27 antibody. Image from verified customer review.
Immunocytochemistry/ Immunofluorescence: HSP27 Antibody - BSA Free [NBP1-75477]

Immunocytochemistry/ Immunofluorescence: HSP27 Antibody - BSA Free [NBP1-75477]

Immunocytochemistry/Immunofluorescence: HSP27 Antibody [NBP1-75477] - Hsp27 Antibody [NBP1-75477] - HSP27 antibody was tested in HeLa cells with FITC (green). Nuclei and actin were counterstained with DAPI (blue) and Phalloidin (red).
Immunohistochemistry: HSP27 Antibody - BSA Free [NBP1-75477]

Immunohistochemistry: HSP27 Antibody - BSA Free [NBP1-75477]

Immunohistochemistry: Hsp27 Antibody [NBP1-75477] - Staining of HSP27 in mouse heart using DAB with hematoxylin counterstain.
Western Blot: HSP27 AntibodyBSA Free [NBP1-75477]

Western Blot: HSP27 AntibodyBSA Free [NBP1-75477]

Western Blot: Hsp27 Antibody [NBP1-75477] - Analysis of HSP27 in human skeletal muscle lysate.

Applications for HSP27 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:100

Immunohistochemistry

1:100

Immunohistochemistry-Paraffin

1:100

Western Blot

2 ug/ml
Application Notes
This HSP27 antibody is useful for ICC/IF, IHC-P, and Western blot where a band is seen ~27 kDa. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.

Reviewed Applications

Read 1 review rated 3 using NBP1-75477 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS and 30% Glycerol

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: HSP27

Heat shock protein 27 is a molecular chaperone involved in protection of cells during cellular stress events. The activity of HSP27 protects from apoptotic events caused by heat shock and other distress. Immunohistochemical detection of HSP27 has been suggested as a qualitative biomarker for colorectal cancer. It is also highly expressed in lung cancer, cerebral inflammation, ischemia-reperfusion injury, and glaucoma. Interestingly, HSP27 overexpression can counteract the activity of multi-drug resistance proteins in breast cancer. Polyphenols downregulate the expression of HSP27, as well as many antioxidant enzymes, resulting in susceptibility to stress and damage.

Long Name

Heat Shock Protein 27

Alternate Names

HSP25, HSPB1

Entrez Gene IDs

3315 (Human); 15507 (Mouse)

Gene Symbol

HSPB1

UniProt

P04792, P14602 (Mouse)

Additional HSP27 Products

Product Documents for HSP27 Antibody - BSA Free

Certificate of Analysis

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Product Specific Notices for HSP27 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for HSP27 Antibody - BSA Free

Customer Reviews for HSP27 Antibody - BSA Free (1)

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  • Name: Anonymous
    Application: Western Blot
    Sample Tested: Whole cell lysate from Jurkat cells, in lysis buffer
    Species: Human
    Verified Customer | Posted 10/07/2014
    HSP27 western in Jurkat cells
    HSP27 Antibody - BSA Free NBP1-75477

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Protocols

View specific protocols for HSP27 Antibody - BSA Free (NBP1-75477):

HSP27 Antibody:
Immunocytochemistry Protocol

Culture cells to appropriate density in 35 mm culture dishes or 6-well plates.

1. Remove culture medium and add 10% formalin to the dish. Fix at room temperature for 30 minutes.
2. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
3. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
4. To block nonspecific antibody binding incubate in 10% normal goat serum from 1 hour to overnight at room temperature.
5. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
6. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
7. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
8. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
9. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

HSP27 Antibody:
Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.

HSP27 Antibody:
Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.

*Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

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